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首页> 外文期刊>Journal of Clinical Microbiology >Use of Pulsed-Field Gel Electrophoresis for Molecular Epidemiologic and Population Genetic Studies ofMycobacterium tuberculosis
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Use of Pulsed-Field Gel Electrophoresis for Molecular Epidemiologic and Population Genetic Studies ofMycobacterium tuberculosis

机译：脉冲场凝胶电泳在结核分枝杆菌的分子流行病学和群体遗传研究中的应用

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Pulsed-field gel electrophoresis (PFGE) is a powerful molecular biology technique which has provided important insights into the epidemiology and population biology of many pathogens. However, few studies have used PFGE for the molecular epidemiology ofMycobacterium tuberculosis. A laboratory protocol was developed to determine the typeability, stability, and reproducibility of PFGE typing of M. tuberculosis. Formal data-analytical techniques were used to assess the genetic diversity elucidated by PFGE analyses using four separate restriction enzymes and by IS6110 RFLP analyses, as well as to assess the concordance among these typing methods. One hundred epidemiologically characterized clinical isolates of M. tuberculosis were genotyped with four different PFGE enzymes (AseI, DraI,SpeI, and XbaI), as well as by RFLP analysis with IS6110. Identical patterns were found among 34 isolates known to be genetically related, suggesting that the PFGE protocol is robust and reproducible. Among 66 isolates representing population-sampled cases, heterozygosity and information content dependency estimates indicate that all five genotyping systems capture quantitatively similar levels of genetic diversity. Nevertheless, comparisons between PFGE analyses and IS6110 typing reveals that PFGE provided more discrimination among isolates with fewer than five copies of IS6110 and less clustering in isolates with five or more copies. The comparisons confirm the hypothesis that the resolution of IS6110 RFLP genotyping is dependent upon the number of IS6110 elements in the genome of isolates. The general concordance among the results obtained with four independent enzymes suggests that M. tuberculosis is a clonal organism. The availability of a robust genotyping technique largely independent of repetitive elements has implications for the molecular epidemiology of M. tuberculosis.
机译：脉冲场凝胶电泳（PFGE）是一种强大的分子生物学技术，它为许多病原体的流行病学和群体生物学提供了重要的见识。然而，很少有研究将PFGE用于结核分枝杆菌的分子流行病学研究。开发了实验室协议，以确定 M的PFGE分型的可打字性，稳定性和可重复性。结核病。正式的数据分析技术被用来评估通过使用四个独立的限制性酶的PFGE分析和IS 6110 RFLP分析所阐明的遗传多样性，以及评估这些分型方法之间的一致性。一百个流行病学特征的临床分离株。用四种不同的PFGE酶（ Ase I， Dra I， Spe I和 Xba ）对结核病进行基因分型em> I），以及使用IS 6110 进行RFLP分析。在已知与遗传相关的34个分离株中发现了相同的模式，这表明PFGE方案是可靠且可重复的。在代表人群抽样病例的66个分离株中，杂合性和信息含量依赖性估计表明，所有五个基因分型系统都在数量上捕获了相似程度的遗传多样性。尽管如此，PFGE分析和IS 6110 分型的比较显示，PFGE在IS 6110 少于五个拷贝的分离株中提供了更多的区分度，而在五个或更多拷贝的分离株中提供了较少的聚类。 。比较结果证实了以下假设：IS 6110 RFLP基因分型的分辨率取决于分离株基因组中IS 6110 元素的数量。用四种独立酶获得的结果之间的总体一致性表明， M。结核病是一种克隆生物。强大的基因分型技术的可用性在很大程度上与重复元素无关，这对 M的分子流行病学有影响。结核病。

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